KMID : 0620920080400020208
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Experimental & Molecular Medicine 2008 Volume.40 No. 2 p.208 ~ p.219
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Transcriptional profiling in human HaCaT keratinocytes in response to kaempferol and identification of potential transcription factors for regulating differential gene expression
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Kang Byung-Young
Lee Yong-Sung Kim Su-Jong Lee Ki-Hwan Hong Il Lee Mi-Ock Min Dae-Jin Chang Ih-Seop Hwang Jae-Sung Park Jun-Seong Kim Duck-Hee Kim Byung-Gee
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Abstract
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Kaempferol is the major flavonol in green tea and exhibits many biomedically useful properties such as antioxidative, cytoprotective and anti-apoptotic activities. To elucidate its effects on the skin, we investigated the transcriptional profiles of kaempferol-treated HaCaT cells using cDNA microarray analysis and identified 147 transcripts that exhibited significant changes in expression. Of these, 18 were up-regulated and 129 were down-regulated. These transcripts were then classified into 12 categories according to their functional roles: cell adhesion/cytoskeleton, cell cycle, redox homeostasis, immune/defense responses, metabolism, protein biosynthesis/modification, intracellular transport, RNA processing, DNA modification/ replication, regulation of transcription, signal transduction and transport. We then analyzed the promoter sequences of differentially-regulated genes and identified over-represented regulatory sites and candidate transcription factors (TFs) for gene regulation by kaempferol. These included c-REL, SAP-1, Ahr-ARNT, Nrf-2, Elk-1, SPI-B, NF-¥êB and p65. In addition, we validated the microarray results and promoter analyses using conventional methods such as real-time PCR and ELISA-based transcription factor assay. Our microarray analysis has provided useful information for determining the genetic regulatory network affected by kaempferol, and this approach will be useful for elucidating gene-phytochemical interactions.
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KEYWORD
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keratinocytes, kaempferol, NF-¥êB, oligonucleotide array sequence analysis, peroxisome proliferator-activated receptors, transcription factors
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